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Biomedical subjects

D N Foster

Publications and source records attributed to D N Foster.

At least 55 records · Page 3Linked to original sources

Tissue-specific alternative splicing of turkey preprovasoactive intestinal peptide messenger ribonucleic acid, its regulation, and correlation with prolactin secretion.

Although vasoactive intestinal peptide (VIP) is a well characterized physiological PRL-releasing factor in avian species, its regulated expression is not fully understood. We cloned complementary DNAs encoding the prepro-turkey VIP (prepro-tVIP) molecule from an adult turkey hypothalamic complementary DNA library. When the amino acid sequence of the prepro-tVIP was compared to chicken and mammalian sequence, it was found that the isolated tVIP molecules lacked the 27-amino acid peptide histidine isoleucine (PHI) portion of the precursor protein. Several tissues showed an alternatively spliced tVIP transcript that lacked the PHI sequence. Only in the hypothalamus did tVIP-specific primer pairs and reverse transcription-polymerase chain reaction produce two alternatively spliced fragments. The larger hypothalamus-specific fragment was subjected to nucleotide sequence analysis and identified as containing the alternatively spliced PHI-containing exon, which encoded a 27-amino acid PHI peptide in addition to the 8 amino acids that flanked the peptide. Hypothalamic tVIP expression was shown to be up-regulated during the incubation phase of the reproductive cycle. The increased steady state level of tVIP messenger RNA appears to be regulated by nesting behavior, because nest deprivation dramatically suppressed its expression. Levels of the minor tVIP transcript containing both the PHI- and VIP-encoding exons did not significantly change between reproductive stages and were maintained at approximately 4-6% of the total tVIP transcript level. Our findings provide further evidence that VIP is the most important PRL-releasing factor in birds. Our study should serve as a useful model for determining whether PHI contributes in any way to the physiological role of PRL regulation. Revealing the tissue distribution of VIP and PHI gene expression and tissue-specific alternative splicing could contribute to an understanding of the physiological functions of the two peptides as well as their relative roles in PRL regulation.

Alternative Splicing↗

Sequence analysis of the turkey LH beta subunit and its regulation by gonadotrophin-releasing hormone and prolactin in cultured pituitary cells.

cDNAs encoding the precursor molecule of the turkey LH beta subunit (tLH beta) were cloned from a turkey pituitary cDNA library. The nucleotide sequence of the longest of two different tLH beta cDNA clones contained 592 bp, and included 23 bp of the 5' untranslated region (UTR) and 92 bp of the 3' UTR in addition to a 477 bp open reading frame that encoded a 39 amino acid leader polypeptide and a 120 amino acid mature apoprotein. Turkey and chicken LH beta sequences shared approximately 92 and 93% nucleotide and amino acid sequence similarities respectively. Northern blot analysis of total cellular anterior pituitary RNA showed that an approximate 800 base transcript hybridized to a 32P-labelled tLH beta cDNA probe. The gonadotrophin-releasing hormone (GnRH)- and prolactin (PRL)-regulated expression of LH and PRL in dispersed pituitary cells was determined by Northern blot analysis of tLH beta and PRL steady-state mRNA levels and by RIA analysis of secreted LH and PRL. GnRH-treated cells showed increased levels of both tLH beta mRNA and secreted LH, whereas mRNA and secreted levels of PRL did not change significantly. Cells treated with PRL showed lower levels of tLH beta and PRL mRNA as well as decreased release of LH and PRL. When cells were treated with both PRL and GnRH, increases in tLH beta mRNA and secreted levels of LH observed with GnRH alone were negated, whereas the decreases in mRNA and secreted levels of PRL observed with PRL alone were abrogated. These findings suggest that PRL can down-regulate tLH beta gene expression and spontaneous release of LH as well as autoregulate PRL gene expression and spontaneous release of PRL, while GnRH appears capable of modulating the effects of PRL-regulated LH and PRL gene expression and spontaneous release.

Amino Acid Sequence↗

Role of vasoactive intestinal peptide in the control of prolactin-induced turkey incubation behavior. I. Acute infusion of vasoactive intestinal peptide.

Vasoactive intestinal peptide (VIP) stimulates prolactin (PRL) secretion. Ovine PRL induces incubation behavior in avian species. This study was designed to determine whether VIP can elevate plasma PRL for up to 3 h. Saline or porcine VIP (pVIP; 30, 60, or 150 ng/min) was infused into the median eminence of laying turkeys for 1 h. The 60- and 150-ng doses of pVIP increased plasma PRL (p < 0.01), whereas the 30-ng dose was insignificant. Pituitary PRL content decreased in pVIP-treated turkeys. Two-hour infusion of 60 or 150 ng chicken VIP (cVIP)/min produced similar elevations of plasma PRL (p < 0.001), which declined within 80 min. Both treatments induced insignificant increases in pituitary PRL mRNA. Saline or cVIP (30, 60, or 60 [pulsed] ng/min) was infused into the median eminence for 3 h. Sixty ng cVIP/min induced the largest PRL release (p < 0.05). The pulsatile and low-cVIP treatments resulted in release of a significant amount of PRL in comparison to the saline treatment (p < 0.01). All cVIP treatments resulted in decreased pituitary PRL content (p < 0.05). The 60-ng dose increased PRL mRNA (p < 0.1). This study shows that 60 ng VIP/min causes the maximum PRL release in laying turkeys. However, pituitary PRL content is depleted and PRL synthesis cannot maintain PRL secretion at high levels.

Animals↗

Role of vasoactive intestinal peptide in the control of prolactin-induced turkey incubation behavior. II. Chronic infusion of vasoactive intestinal peptide.

Hyperprolactinemia is associated with incubation behavior in avian species. Increased nesting activity is a major indication of incubation behavior. Vasoactive intestinal peptide (VIP) stimulates prolactin (PRL) secretion from the anterior pituitary. The goal of this study was to induce incubation behavior by stimulating PRL through chronically infusing VIP into the third ventricle of turkey brains. In experiment 1, porcine VIP (pVIP) was infused into the median eminence at a rate of 60 ng/min for 7 days by means of osmotic pumps implanted s.c.. Plasma PRL increased significantly in the pVIP-treated turkeys (p < 0.001). Although egg laying was not affected by the pVIP infusion, the mean oviduct weight decreased (p < 0.057). In experiment 2, saline or pVIP (30 or 60 ng/min) was infused into the third ventricle of laying turkeys for 12 days. Both pVIP treatments increased plasma PRL for 9 days (p < 0.05). The 30-ng pVIP/min infusion decreased nesting activity, plasma LH, ovary and oviduct weight, hypothalamic GnRH I, and anterior pituitary VIP receptors (p < 0.1). However, ovine PRL infusion (20.8 ng/min) into the same turkey flock increased nesting activity (p < 0.01). In conclusion, pVIP does not induce incubation behavior in laying turkeys.

Animals↗

Theca cell cytochrome P450 17-hydroxylase and aromatase messenger ribonucleic acid abundance and serum steroid levels during follicular atresia associated with incubation behavior in the domestic turkey hen.

This study was designed to examine changes in cytochrome P450 17 alpha-hydroxylase (C17) and aromatase (ARO) mRNA contents in the theca layer of preovulatory follicles (POF) as turkey hens transit from egg laying to incubation. Hens were grouped into the following categories: 1) laying hens--laid one egg per day and nested 1-2 times per day; 2) transitional hens--laid one egg per day and nested > 4 times per day; and 3) Day 1, Day 3, and Day 5 incubating hens--laid no eggs for 2, 4, or 6 days, respectively, and nested > 4 times per day. Small white follicles (SWF) and the theca layer from the largest (F1) and the third (F3), fifth (F5), and seventh (F7) largest POF were dispersed and challenged with testosterone (T) for 5 h. Relative levels of C17 and ARO mRNA were examined from the theca layers of F1, F3, F5, F7, and SWF. The number of atretic follicles increased from 0 (layers) to 8 (Day 5 incubating hens). Serum LH, progesterone (P), and estradiol (E), but not T, declined on Day 1 of incubation. Basal levels of P, T, and E from theca and SWF cells declined in incubating hens. Both basal and T-stimulated theca and SWF production of E decreased in incubating hens. C17 and ARO mRNA declined in SWF, F7, and F5 during follicular atresia. It is suggested that reduced gene expression of ovarian steroidogenic enzymes may be a partial determinant of reduced circulating sex steroid levels in incubating hens.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Pelvic stabilization during resistance training: its effect on the development of lumbar extension strength.

The purpose of this study was to evaluate and compare resistance exercise training with and without pelvic stabilization on the development of isolated lumbar extension strength. Isometric torque of the isolated lumbar extensor muscles was measured at seven positions through a 72 degree range-of-motion on 47 men and 30 women before and after 12 weeks of variable resistance lumbar extension training. Subjects were assigned to either a group that trained with pelvic stabilization (P-STAB, n = 21), a group that trained without pelvic stabilization (NO-STAB, n = 41), or a control group that did not train (n = 15). Subjects trained once a week with 8 to 12 repetitions to volitional exhaustion. The P-STAB and NO-STAB groups showed significant (p < or = 0.05) and similar increases in the weight load used for training (P-STAB = 24.1 +/- 9.4kg; NO-STAB = 19.4 +/- 11.0kg) during the 12-week training period. In contrast, posttraining isometric torque values describing isolated lumbar extension strength improved only for the P-STAB group (23.5%, p < or = 0.05) and not for the NO-STAB group (-1.2%, p > 0.05) relative to controls. These data indicate that pelvic stabilization is required to effectively train the lumbar extensor muscles. The increased training load for the NO-STAB group is probably the result of exercising the muscles involved in pelvic rotation (hamstring and buttock muscles).

Adult↗

Ovarian involvement in the suppression of luteinizing hormone in the incubating turkey (Meleagris gallopavo).

A series of experiments were conducted to elucidate the role of the ovary in incubation behavior-associated luteinizing hormone (LH) suppression. Ovariectomy (Ovx) increased (p < 0.05) serum LH levels in nonphotostimulated, laying and photorefractory turkeys but not in incubating birds (p > 0.05). Ovx had no effect on intramuscularly injected mammalian luteinizing hormone-releasing hormone (4 micrograms/kg i.m.)-induced LH in incubating hens, but enhanced the LH response in laying hens. Serum LH and prolactin (Prl) were unaffected by Ovx, and nest deprivation (ND) decreased Prl levels (p < 0.05) with no effect on serum LH of incubating turkeys. However, serum LH increased (p < 0.05) and Prl decreased in Ovx-ND birds. Prl mRNA abundance (11.9 +/- 1.2 ng/microgram total RNA) decreased following Ovx (3.4 +/- 0.4 ng/microgram total RNA) or ND (3.6 +/- 0.5 ng/microgram total RNA). Nest deprivation increased LH beta mRNA (2.5-fold) which was further increased (4.8-fold) by Ovx. Hypothalamic GnRH-I and GnRH-II contents increased (p < 0.05) in Ovx-ND turkeys. We conclude that serum LH suppression during incubation behavior requires ovarian participation acting synergistically with elevated Prl and/or nesting stimulus on hypothalamic GnRH, and that the concentration fo LH beta mRNA may be a limiting factor in LH secretion.

Animals↗

Ontogeny of pituitary growth hormone and growth hormone mRNA in the chicken.

The changes in pituitary growth hormone (GH) mRNA levels have been determined by Northern blot analysis and laser densitometry during embryonic development and posthatch growth of white Leghorn cockerels. Pituitary GH mRNA levels were observed to progressively increase between 18 days of embryonic development to a maximum at 4 weeks of age (posthatch). Subsequently, pituitary GH mRNA levels declined between 4 and 8 weeks of age, and between 12 weeks of age and adulthood. Pituitary GH contents showed increases during embryonic development and posthatch growth that paralleled the rise in GH mRNA. The decline in pituitary GH mRNA levels between 4 weeks of age and adulthood occurs when GH secretion has been observed previously to decline.

Aging↗

Positive and negative cis-acting regulatory elements mediate expression of the mouse vascular smooth muscle alpha-actin gene.

Segments of the 5'-flanking region of the mouse vascular smooth muscle alpha-actin gene were assayed for promoter activity in transfected mouse BC3H1 myogenic cells and AKR-2B embryonic fibroblasts. The region between -150 and -191 that functions as a positive transcriptional element in myogenic and fibroblastic cells contains a mammalian-specific inverted CC(A/T)6GG-type consensus sequence. Expression was restricted to fully differentiated myogenic cells when an additional sequence spanning -191 to -224 was included in reporter gene constructs. This 33-base pair (bp) negative regulatory element is 70% conserved between the mouse and human genes and contains a 10-bp motif at its 3' end that only partially resembles a CC(A/T)6GG element. Retention of a GGGA motif at the 3' boundary of the 33-bp region is sufficient to maintain full transcriptional repression in fibroblasts and is partly responsible for repression in undifferentiated myoblasts. Complete muscle tissue-restrictive expression requires an additional 8 bp from the CC(A/T)6GG-like element immediately 5' to the GGGA motif, since replacement of this region with an unrelated 10-bp sequence completely eliminated restrictive transcriptional behavior in undifferentiated myoblasts. The distal portion of the 5'-flanking region between -224 and -1074 contains six E-box motifs (CANNTG) and mediates high level transcription only in postconfluent BC3H1 myoblasts. Analysis of reporter gene constructs including either the proximal E-box at -240 or all six E-boxes indicate that the five distal E-boxes are not required for high level transcription. A 724-bp segment of the 5'-flanking region consisting of the proximal E-box flanked upstream by a mammalian-specific 352-bp region was sufficient for maximal transcriptional activation in postconfluent BC3H1 myoblasts. Deletion of the 352-bp region restricts the early transcriptional response to high cell density in temporal studies of promoter activity during BC3H1 myogenic cell differentiation.

Actins↗

Density-dependent modulation of vascular smooth muscle alpha-actin biosynthetic processing in differentiated BC3H1 myogenic cells.

The expression of vascular smooth muscle (VSM) alpha-actin mRNA during BC3H1 myogenic cell differentiation is specifically stimulated by conditions of high cell density. Non-proteolytic dissociation of cell-cell and cell-matrix contacts in post-confluent cultures of BC3H1 myocytes using EDTA promotes loss of the differentiated morphological phenotype. EDTA-dispersed myocytes exhibit an undifferentiated fibroblastoid appearance and contained reduced levels of both VSM and skeletal alpha-actin mRNA. Muscle alpha-actin mRNA levels in EDTA-dispersed myocytes were not restored to that observed in confluent myocyte preparations by experimental manipulation of cell density conditions. Pulse-labeling techniques using L-[35S]cysteine to identify muscle actin biosynthetic intermediates revealed that EDTA-dispersed myocytes expressed nascent forms of both the VSM and skeletal muscle alpha-actin polypeptide chains. However EDTA-dispersed myocytes were less efficient in the post-translational processing of immature VSM alpha-actin compared to non-dispersed myocytes. Simple cell-to-cell contact may mediate VSM alpha-actin processing efficiency since high-density preparations of EDTA-dispersed myocytes processed more VSM alpha-actin intermediate than myocytes plated at low density. The actin isoform selectivity of the response to modulation of intercellular contacts suggests that actin biosynthesis in BC3H1 myogenic cells involves mechanisms capable of discriminating between different isoform classes of nascent actin polypeptide chains.

Actins↗

Activation of a muscle-specific actin gene promoter in serum-stimulated fibroblasts.

Treatment of AKR-2B mouse fibroblasts with serum growth factors or inhibitors of protein synthesis, such as cycloheximide, results in a stimulation of cytoskeletal beta-actin transcription but has no effect on transcription of muscle-specific isotypes, such as the vascular smooth muscle (VSM) alpha-actin gene. Deletion mapping and site-specific mutagenesis studies demonstrated that a single "CArG" element of the general form CC(A/T)6GG was necessary and possibly sufficient to impart serum and cycloheximide-inducibility to the beta-actin promoter. Although the VSM alpha-actin promoter exhibits at least three similar sequence elements, it remained refractory to serum and cycloheximide induction. However, deletion of a 33 base pair sequence between -191 and -224 relative to the transcription start site resulted in the transcriptional activation of this muscle-specific promoter in rapidly growing or serum-stimulated fibroblasts. Although the activity of this truncated promoter was potentiated by cycloheximide in a manner indistinguishable from that of the beta-actin promoter, this was dependent on a more complex array of interacting elements. These included at least one CArG box and a putative upstream activating element closely associated with the -191 to -224 inhibitory sequences. These results demonstrate that the expression of a muscle-specific actin gene in fibroblasts is suppressed by a cis-acting negative control element and that in the absence of this element, the promoter is responsive to growth factor-induced signal transduction pathways.

Actins↗

Ca2+ channel modulation and kinase-C activation in a pituitary cell line: induction of immediate early genes and inhibition of proliferation.

We have studied the interaction between dihydropyridine (DHP) Ca2+ modulators and the phorbol ester phorbol 12-myristate 13-acetate (PMA) on whole cell Ca2+ currents, 45Ca2+ uptake, immediate early gene (IEG) expression, and proliferation in the rat pituitary GH4C1 cell line. When short (3- to 5-msec) depolarizing voltage clamp steps were used to activate L-type Ca2+ channels, the DHP Ca2+ agonist (-)Bay K 8644 markedly enhanced Ca2+ entry by slowing channel closing upon repolarization. In contrast, the Ca2+ agonist induced only small and inconsistent increases in c-fos mRNA and did not measurably increase NGFI-A. Ca2+ channel activation by depolarization with 50 mM KCl in the presence of (-)Bay K 8644 induced large increases in 45Ca2+ uptake, but failed to markedly induce either of the IEGs. The phorbol ester PMA did not alter T- or L-type Ca2+ current or 45Ca2+ uptake by GH4C1 cells, but triggered large increases in both c-fos and NGFI-A mRNA. In combination, PMA and (-)Bay K 8644 acted synergistically to increase mRNAs for both IEGs. The effect of the DHPs was stereospecific; (+)Bay K 8644, a Ca2+ antagonist, inhibited PMA-induced increases in c-fos and NGFI-A mRNAs. Both PMA and (-)Bay K 8644 inhibited the proliferation of GH4C1 cells, measured by cell count or [3H]thymidine incorporation. The inhibition by the Ca2+ agonist was stereoselective and approximately additive to that of PMA. These results indicate that the expression of c-fos IEG and that of NGFI-A IEG are differentially regulated by separate second messenger pathways in GH4C1 cells.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Nucleotide sequence of the cDNA encoding the common alpha subunit of the chicken pituitary glycoprotein hormones.

Recombinant cDNA clones that encode the alpha subunit of the chicken pituitary glycoprotein hormones were isolated from a pituitary library. The longer of the two cDNA clones that were sequenced was 754 bp in length. It contained 81 nucleotides of the 5'-untranslated region (UTR), an open-reading frame of 360 bp that encoded a 24 amino acid leader polypeptide sequence as well as the 96 amino acid mature alpha subunit, and 268 nucleotides of the 3'-UTR, followed by a 45 bp poly(A) tract. There was 69-79% homology between the nucleotide sequence of the coding region for the chicken and mammalian alpha-subunit cDNAs. Northern blot analysis revealed that the steady-state levels of an approximately 800 bp alpha-subunit specific transcript increased quantitatively when dispersed chicken pituitary glands were treated in culture with chicken gonadotrophin-releasing hormone-I.

Amino Acid Sequence↗

Restoration of juvenile baseline growth hormone secretion with preservation of the ultradian growth hormone rhythm by continuous delivery of growth hormone-releasing factor.

The ability of continuously delivered GH-releasing factor (GRF) to enhance GH secretion while maintaining the normal ultradian GH rhythm was investigated. Synthetic human GH-releasing factor (hGRF(1-44)NH2) was continuously infused for 4 days by means of i.v. catheters to 11-week-old broiler chickens. At this age, overall endogenous GH secretion is low, and baseline GH is barely detectable. Six birds per treatment received vehicle (control), 0.324 mg hGRF(1-44)NH2/kg body weight per day (low dose) or 3.24 mg hGRF(1-44)NH2/kg body weight per day (high dose). After 4 days of GRF conditioning, concurrent with continued GRF infusion, serial blood samples were removed via atrial catheters at 15-min intervals for 6 h and GH plasma profiles determined. High dose GRF significantly increased GH plasma concentrations over tenfold compared with controls; however, most of this increase reflected an increase in basal GH, which was reinstated to juvenile baseline levels. Augmentation of pulse amplitude above this increased baseline was not proportionately as high, and failed to reach juvenile levels. The ultradian rhythm of GH was not altered by continuous GRF administration. Both low and high dose GRF treatments resulted in significant enlargement of the anterior pituitary gland. Total pituitary GH mRNA levels, although elevated over twofold by GRF treatment, were not significantly different from controls. Measures of plasma GH magnitude (overall and baseline mean, and peak amplitude) were significantly correlated with pituitary GH mRNA for control birds, but were not correlated for GRF treatments. Feed intake was markedly depressed (33%) on the high dose GRF treatment, in conjunction with total inhibition of body weight gain over the 4-day period of administration. Longitudinal bone growth and width of the epiphyseal growth plate were also significantly reduced by high dose GRF treatment, probably reflecting the reduced level of nutrient intake, despite high circulating concentrations of GH.

Animals↗

The turkey major histocompatibility complex: identification of class II genotypes by restriction fragment length polymorphism analysis of deoxyribonucleic acid.

Using a chicken Class II MHC clone in Northern blot analysis, tissue-specific expression of turkey Class II MHC genes was observed in the embryonic bursa of Fabricius as well as in the adult spleen. In contrast, there was no detectable expression in the embryonic liver, brain, or spleen. Southern blot analysis of BamHI-digested turkey DNA revealed two restriction fragment length polymorphism (RFLP) patterns that did not deviate significantly from single-gene Mendelian inheritance. Further analysis of PvuII-digested DNA from 325 turkeys showed four distinct RFLP patterns that segregated within the turkey lines studied. Because the chicken Class II MHC clone hybridized specifically to mRNA in immune-associated tissues, and because it identified polymorphisms among turkeys, the chicken clone is suggested to identify four turkey Class II MHC genotypes. The current study provides good evidence that RFLP analysis of DNA can be used as a means for molecular genotyping at the MHC in turkeys.

Animals↗

Pattern of secretion of luteinizing hormone and testosterone in the sexually mature male turkey.

Whether luteinizing hormone (LH) and testosterone (T) are secreted in pulsatile patterns was determined in sexually mature male turkeys. Turkeys were chronically cannulated and serially bled for three 8-hr periods covering the 24-hr day (14L:10D, n = 7, series B), or for two 12-hr periods covering the 24-hr day (14L:10D, n = 4, series C). Pulses of both LH and T occurred during both the light and dark portions of the 24-hr day. A portion of the secretory episodes of T, where the baseline level of LH was relatively low, was associated with prior peaks of LH secretion. Secretory episodes of T also occurred, where baseline levels of LH and T were both relatively high, without detection of prior peaks of LH. No differences were found between the photophase and scotophase portions of the photoperiod for either LH or T concentration. It is concluded that T is secreted in a pulsatile pattern in sexually mature male turkeys. However, LH is secreted in a pulsatile pattern only when baseline levels of both LH and T are relatively low. Neither LH nor T secretion is entrained by the photoperiod. Corticosterone was measured in hourly samples, but no changes in concentration occurred in association with the photoperiod.

Animals↗

Symptomatic pancreatic heterotopia treated by local excision.

Non-ulcer dyspepsia is a continuing problem and in many cases a precise cause is never identified. We present five patients with an allegedly uncommon condition--pancreatic heterotopia. They were managed by local excision of the tumour and after a mean (range) follow up of 42 (9-80) months all remain free of the original symptoms.

Adult↗